BACKGROUND OF THE INVENTION
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1. Field of the Invention [0001]
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The present invention relates to a novel gene. In particular, the present invention relates to a novel gene in plants which encodes a protein having the function of controlling an in-vivo signal transduction system in a physiological reaction system against brassinosteroid hormone. [0002]
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2. Description of the Related Art [0003]
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Transposons are mutagenic genes which are known to be ubiquitous in animal, yeast, bacterial, and plant genomes. Transposons are classified into two classes, Class I and Class II, depending on their transposition mechanisms. Transposons belonging to Class II are transposed in the form of DNAs without being replicated. Known Class II transposons include the Ac/Ds, Spm/dSpm and Mu elements of [0004] Zea mays (Fedoroff, 1989, Cell 56, 181-191: Fedoroff et al., 1983, Cell 35, 235-242; Schiefelbein et al., 1985, Proc. Natl. Acad. Sci. USA 82, 4783-4787), and the Tam element of Antirrhinum majus (Bonas et al., 1984, EMBO J., 3, 1015-1019). Class II transposons are widely used for gene isolation techniques which utilize transposon tagging. Such techniques utilize the fact that a transposon induces physiological and morphological changes when inserted into genes. The affected gene can be isolated by detecting such changes (Bancroft et al., 1993, The Plant Cell, 5, 631-638; Colasanti et al., 1998, Cell, 93, 593-603: Gray et al., 1997, Cell, 89, 25-31; Keddie et al., 1998, The Plant Cell, 10, 877-887: Whitham et al., 1994, Cell, 78, 1101-1115).
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Transposons belonging to Class I, also referred to as retrotransposons, are replicated and transposed via RNA intermediates. Class I transposons were first identified and characterized in Drosophila and in yeasts. However, recent studies have revealed that Class I transposons are ubiquitous in plant genomes and account for a substantial portion of the genomes (Bennetzen, 1996, Trends Microbiolo., 4, 347-353; Voytas, 1996, Science, 274, 737-738). A large majority of retrotransposons appear to be inactive. Recent studies indicate that some of these retrotransposons are activated under stress conditions such as injuries, pathogenic attacks, or cell culture (Grandbastien, 1998, Trends in Plant Science, 3, 181-187; Wessler, 1996, Curr. Biol. 6, 959-961; Wessler et al., 1995, Curr. Opin. Genet. Devel. 5, 814-821). Activation under stress conditions has been reported for Tnt1 A and Tto1 in tobacco (Pouteau et al., 1994, Plant J., 5, 535-542: Takeda et al., 1988, Plant Mol. Biol., 36, 365-376), and Tos17 in rice (Hirochika et al., 1996, Proc. Natl. Acad. Sci. USA, 93, 7783-7788), for example. [0005]
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The Tos17 retrotransposon of rice is one of the most-extensively studied plant Class I elements in plants. Tosl7 was cloned by an RT-PCR method using a degenerate primer prepared based on a conservative amino acid sequence in reverse transcription enzyme domains between Ty1-copia retroelements (Hirochika et al., 1992, Mol. Gen. Genet., 233, 209-216). Tos17 is 4.3 kb long, and has two 138 bp LTRs (long chain terminal repetitions) and PBS (primer binding sites) complementary to the 3′ end of the start methionine tRNA (Hirochika et al., 1996, supra). Tos17 transcription is strongly activated through tissue culture, and its copy number increases with culture time. In Nipponbare, a model Japonica cultivar used for genome analysis, two copies of Tos17 are initially present, which are increased to 5 to 30 copies in a regenerated plant after tissue culture (Hirochika et al., 1996, supra). Unlike Class II transposons which were characterized in yeasts and Drosophila, Tos17 is transposed in chromosomes in random manners and causes stable mutation, and therefore provides a powerful tool for functional analysis of rice genes (Hirochika, 1997, Plant Mol. Biol. 35, 231-240; 1999, Molecular Biology of. Rice (ed. by K. Shimamoto, Springer-Verlag, 43-58). [0006]
SUMMARY OF THE INVENTION
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The present invention relates to a polynucleotide encoding a plant gene capable of controlling a signal transduction system for brassinosteroid hormone, the polynucleotide encoding an amino acid sequence from Met at [0007] position 1 to Arg at position 1057 of SEQ ID NO: 2 in the SEQUENCE LISTING, including any polynucleotide encoding an amino acid sequence in which one or more amino acids are deleted, substituted or added to the amino acid sequence.
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In one embodiment of the invention, the polynucleotide may be derived from rice. [0008]
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In another embodiment of the invention, the polynucleotide may be as represented by SEQ ID NO: 1 in the SEQUENCE LISTING. [0009]
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The present invention further relates to methods for controlling various effects in plants in which brassinosteroid hormone is involved, e.g., growth promotion, yield increase, quality improvement, maturation enhancement, and tolerance against biotic and abiotic stresses. [0010]
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The inventors diligently conducted systematic analyses of phenotypes of plants having a newly transposed To1 7 copy and sequences adjoining Tos17 target sites with respect to rice. As a result, the inventors found a dwarf rice mutation obtained from Tos17 insertion, and isolated the gene responsible for this mutation by utilizing Tos17 as a tag, thereby accomplishing the present invention. [0011]
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Thus, the invention described herein makes possible the advantage of providing a novel plant gene which can be provided by using Tos17. [0012]
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This and other advantages of the present invention will become apparent to those skilled in the art upon reading and understanding the following detailed description with reference to the accompanying figures.[0013]
BRIEF DESCRIPTION OF THE DRAWINGS
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FIGS. 1A and 1B are photographs showing a brassinosteroid non-sensitive mutant having Tos17 inserted therein, which was found among regenerated Akitakomachi lineage. On the left of each figure is shown a brassinosteroid non-sensitive mutant having Tos17 inserted therein. On the right of each figure is shown a wild type plant body. FIG. 1A evidences an influence toward dwarfism and upright form. FIG. 1B evidences an influence toward malformation of grain hulls. [0014]
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FIGS. 2A and 2B are photographs showing a brassinosteroid non-sensitive mutant having Tos17 inserted therein, which was found among regenerated Nipponbare lineage. On the left of each figure is shown a brassinosteroid non-sensitive mutant having Tos17 inserted therein. On the right of each figure is shown a wild type plant body. FIG. 2A evidences an influence toward dwarfism and upright form. FIG. 2B evidences an influence toward malformation of grain hulls. [0015]
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FIG. 3A shows a Northern analysis autoradiogram of RNA extracted from the leaves of a brassinosteroid non-sensitive mutant (Akitakomachi) and RNA extracted from various organs of a wild type rice plant (Nipponbare). [0016]
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FIG. 3B shows a Northern analysis autoradiogram of RNA extracted from brassinosteroid non-sensitive mutants and RNA extracted from wild type rice plants. The left-hand side of FIG. 3B shows a comparison between wild types and mutants obtained by using a 5′ probe. The right-hand side of FIG. 3B shows a comparison between wild types and mutants obtained by using a 3′ probe. [0017]
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FIG. 4 shows an amino acid sequence of the novel rice gene which controls a physiological reaction system induced by brassinosteroid hormone, together with characteristic sequences found therein (where nuclear localization signals and an ATP/GTP binding motif can be observed). [0018]
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FIG. 5A shows a brassinosteroid leaf blade bending experimentation using a mutated line (A0369) derived from Akitakomachi. The left-hand side shows results of wild type plants, whereas the right-hand side shows results of mutants. [0019]
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FIG. 5B shows a brassinosteroid leaf blade bending experimentation using a mutated line (NC6148) derived from Nipponbare. The left-hand side shows results of wild type plants, whereas the right-hand side shows results of mutants.[0020]
DESCRIPTION OF THE PREFERRED EMBODIMENTS
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The present invention provides a method for producing an improved plant, the method utilizing a novel plant gene which can be provided by using Tos17. [0021]
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According to the present invention, there is provided a polynucleotide encoding a plant gene capable of controlling various effects in which brassinosteroid hormone is involved. As used herein, the term “capable of controlling various effects” means the ability to control various effects in plants in which brassinosteroid hormone is involved, e.g., growth promotion, yield increase, quality improvement, maturation enhancement, and tolerance against biotic and abiotic stresses, including dwarfism, upright form, and malformation of grain hulls, thereby providing a number of agriculturally useful effects as are attained by treatments with brassinosteroid hormone agricultural chemicals. The term “plants” encompasses both monocotyledons and dicotyledons. [0022]
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A polynucleotide encoding a plant gene capable of controlling a signal transduction system for brassinosteroid hormone according to the present invention is, for example, a polynucleotide encoding an amino acid sequence from Met at [0023] position 1 to Arg at position 1057 of SEQ ID NO: 2 in the SEQUENCE LISTING, including any polynucleotide encoding an amino acid sequence in which one or more amino acids are deleted, substituted or added to the aforementioned amino acid sequence.
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A polynucleotide encoding a plant gene capable of controlling various effects in which brassinosteroid hormone is involved encompasses any polynucleotides which have at least about 80% sequence homology, preferably at least about 85% sequence homology, and more preferably at least about 90% sequence homology, still more preferably at least about 95% sequence homology, and most preferably at least about 99% sequence homology, with an amino acid sequence from Met at [0024] position 1 to Arg at position 1057 of SEQ ID NO: 2 in the SEQUENCE LISTING, so long as they are capable of controlling various effects in plants in which brassinosteroid hormone is involved. The term “sequence homology” indicates a degree of identity between two polynucleotide sequences to be compared with each other. The rate (%) of sequence homology between two polynucleotide sequences for comparison is calculated by, after optimally aligning the two polynucleotide sequences for comparison, obtaining a matched position number indicating the number of positions at which identical (“matched”) nucleic acid bases (e.g., A, T, C, G, U, or I) are present in both sequences, dividing the matched position number by total number of bases in the polynucleotide sequences for comparison, and multiplying the quotient by 100. The sequence homology can be calculated by using the following sequencing tools, for example: a Unix base program designated GCG Wisconsin Package (Program Manual for the Wisconsin Package, Version 8, September 1994, Genetics Computer Group, 575 Science Drive Madison, Wis., USA 53711; Rice, P. (1996) Program Manual for EGCG Package, Peter Rice, The Sanger Centre, Hinxton Hall, Cambridge, CB10 1RQ, England), and the ExPASy World Wide Web molecular biology server (Geneva University Hospital and University of Geneva, Geneva, Switzerland).
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Cells into which genes have been introduced are first selected based on drug resistance, e.g., hygromycin resistance, and then regenerated into plant bodies by using usual methods. [0025]
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The terminology and laboratory procedures described throughout the present specification are directed to those which are well-known and commonly employed in the art. Standard techniques may be used for recombination methods, polynucleotide synthesis, microorganisms culturing, and transformation (e.g., electroporation). Such techniques and procedures are generally known from various standard textbooks available in the field or by way of the present specification (including a generally-referenced textbook by Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd edition (1989) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.). Such literature is incorporated herein by reference. [0026]
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The polynucleotide according to the present invention can be obtained by using the method described herein, for example. However, the polynucleotide according to the present invention may also be obtained by any chemical synthesis process based on the sequence disclosed herein. For example, the polynucleotide according to the present invention may be synthesized by using a polynucleotide synthesizer available from Applied Bio Systems in accordance with the instructions provided by the manufacturer. [0027]
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Methods of PCR amplification are well-known in the art (PCR Technology: Principles and Applications for DNA Amplification, ed. H A Erlich, Freeman Press, NewYork, N.Y. (1992); PCR Protocols: A Guide to Methods and Applications, Innis, Gelfland, Snisky, and White, Academic Press, San Diego, Calif. (1990); Mattila et al. (1991) Nucleic Acids Res. 19: 4967; Eckert, K. A. and Kunkel, T. A. (1991) PCR Methods and Applications 1: 17; PCR, McPherson, Quirkes, and Taylor, IRL Press, Oxford). Such literature is incorporated herein by reference. [0028]
EXAMPLES
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Hereinafter, the present invention will be described by way of examples which are of illustrative but non-limiting nature. [0029]
Example 1
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Activation of Tos17 Via Culture [0030]
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Using fully ripened seeds of Nipponbare and Akitakomachi, which are varieties of Japonica subspecies, induction of calli and cell suspension culture were carried out as described earlier (Hirochika et al., 1996, supra). The activation of Tos17 which was used for gene destruction was carried out following the method of Ohtsuki (1990) (rice protoplast culture system, Food and Agricultural Research Development Association). In summary, fully ripened seeds of rice were cultured in an MS medium having 2,4-dichlorophenoxyacetic acid (2,4-D) added thereto (2 mg/ml) (Ohtsuki (1990), supra) (25° C., 1 month), to induce callus formation. The resultant calluses were cultured for 5 months in an N6 liquid medium having 2,4-D added thereto (Ohtsuki (1990), supra), and thereafter placed on a redifferentiation medium (Ohtsuki (1990), supra), whereby redifferentiated rice plants were obtained (first generation (R1) plants). [0031]
Example 2
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Isolation of sequences adjoining Tos17 [0032]
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Utilizing each of the regenerated R1 rice plants obtained according to Example 1 as a first strain, about 1000 R1 seeds were collected from each strain and grown on a paddy field to obtain second generation (R2) plants, which were subjected to a morphological analysis. As a result of observing the phenotypes of the respective plant bodies in the R2 group, it was learned that about ¼ of the R2 group of an Akitakomachi strain A0369 exhibit the “dwarfism, upright form, and malformation of grain hulls” phenotype (FIGS. 1A and 1B). In the regenerated group of Akitakomachi, dwarfism, upright form, and malformation of grain hulls were observed for brassinosteroid insensitive mutants (FIG. 1A, left, and FIG. 1B, left), as compared with the wild type (FIG. 1A, right, and FIG. 1B, right). The isolation of adjacent sequences of transposed Tos17, which is co-segragating with the phenotypes, was carried out by an IPCR method (Ochman et al., Genetics Nov; 120(3): 621-3(1988) and Triglia et al., Nucleic Acids Res Aug,25: (16): 8186(1988)) The total DNA of A0369 was digested with XbaI, and a ligation process was performed in a large quantity of solution, thereby obtaining self-ligated circular molecules. In the self-ligated circular molecules, the adjacent sequences are flanking the internal sequence of Tos17. As a result, amplification was successfully carried out by usual PCR methods using an outward primer pair (T17TAIL3: GAGAGCATCATCGGTTACATCTTCTC; T17-1950R: TCTAGCAGTCTCAATGATGTGGCG) based on the known sequence of Tos17. [0033]
Example 3
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Search for Alleles [0034]
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Using the sequence obtained according to Example 2, lineage in which Tos17 had been inserted at a different site in the same gene was selected by PCR from the regenerated rice group of Nipponbare. As a result, a line (NC6148) which similarly exhibited dwarfism, upright form, and malformation of grain hulls were observed for brassinosteroid was selected. That is, in the regenerated rice group of Nipponbare, as well, dwarfism, upright form, and malformation of grain hulls were observed for brassinosteroid insensitive mutants (FIG. 2A, left, and FIG. 2B, left), relative to the wild type (FIG. 2A, right, and FIG. 2B, right). It was concluded that these common mutations were results of the same gene having been destroyed. [0035]
Example 4
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Expression and Analysis of the Causative Gene [0036]
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From the group of R2 rice plants (selfed progeny from the A0369 and NC6148 strains) obtained according to Examples 2 and 3, individuals exhibiting mutation were identified from normal individuals. RNA was prepared from both groups of individuals, and the expression specificity was analyzed through Northern analysis. After agarose electrophoresis, the RNA obtained from individuals exhibiting mutation and the RNA obtained from normal individuals were allowed to adsorb to nylon membranes. DNA fragments which were obtained by amplifying via PCR a sequence (positions 5775-6638 of the genomic sequence) on the 5′ side and a sequence (positions 8175-8765 of the genomic sequence) on the 3′ side of the Tos17 insertion site in both mutated lines were labeled with [0037] 32P-dCTP. By using these as probes, expression specificity was analyzed through Northern analysis (FIGS. 3A and 3B). As seen from the Northern analysis autoradiogram shown in FIG. 3A, a band (about 4.3 kb) indicated by an arrow was confirmed to be expressed in all observed organs of the wild type. However, in the mutants, transcripts of abnormal sizes were observed due to the insertion of Tos17, indicating that the natural function of the wild type is lost (FIG. 3B).
Example 5
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Structural Analysis of the Causative Gene [0038]
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Using the sequence obtained according to Example 2 as a probe, the corresponding cDNA and genomic clone were obtained from a cDNA library and a genomic library. Their structures are shown in SEQ ID Nos: 1 and 3. It was learned that this gene includes 6 exons and 5 introns, encoding 1057 amino acids, and that Tos17 had been inserted at the 4th and 5th exons in two mutants, respectively. Moreover, motif search results suggested the presence of nuclear localization signal 1 (amino acid residues 329-367 of SEQ ID NO: 2, Robbins & Dingwall consensus sequence; a search result by PSORT program (http://psort.ims.u-tokyo.ac.jp/)) and nuclear localization signal 2 (amino acid residues 457-460, 595-600 of SEQ ID NO: 2, 4 amino acid nuclear localization pattern signal; a search result by PSORT program (http://psort.ims.u-tokyo.ac.jp/)) as well as the presence of an ATP/GTP binding domain (amino acid residues 526-533 of SEQ ID NO: 2; a search result by a motif search service on Genomenet(http://www.genome.ad.jp/)). Thus, the possibility of this gene being involved in signal transduction was suggested (FIG. 4). [0039]
Example 6
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Brassinosteroid Sensitivity Evaluation [0040]
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The present gene was deduced to be a factor involved in the signal transduction system for plant hormones, taking note of the facts that the present gene was expressed in all plant bodies and that pleiotropic influences resulted from destroying this gene, as well as the possibility that the gene might be a factor involved in the signal transduction system. Presuming that the signal transduction system is that for brassinosteroid hormone in view of the resultant upright form, the inventors performed a leaf blade bend response test as a brassinosteroid response test, by using brassinolide, which is one kind of brassinosteroid hormone. The second leaf of rice which was allowed to germinate in the dark was cut off, and immersed in a 1 ng/ml of brassinosteroid solution for 48 hours. The wild type individuals having the wild type genes showed bending of the leaf blades and leaf sheath junctions (left-hand side in FIGS. 5A and 5B), showing response to brassinolide, whereas mutant individuals showed little bending thereof (right-hand side in FIGS. 5A and 5B), indicating that the destruction of the present gene resulted in the loss of response to brassinosteroid. From the above results, it was revealed that the present gene is a gene involved in the signal transduction system for brassinosteroid hormone. [0041]
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The above examples are illustrative, and by no means limiting, of various aspects of the present invention and the manners in which the oligonucleotide according to the present invention can be made and utilized. [0042]
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Thus, according to the present invention, a novel polynucleotide is provided which is capable of controlling various effects in which brassinosteroid hormone is involved, the polynucleotide being of use in plant breeding. By introducing the present polynucleotide into plants and artificially controlling various effects in which brassinosteroid hormone is involved, it is expected that effects such as growth promotion, yield increase, quality improvement, maturation enhancement, and tolerance against biotic and abiotic stresses can be controlled, thereby providing a number of agriculturally useful effects as are attained by treatments with brassinosteroid hormone agricultural chemicals. [0043]
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Various other modifications will be apparent to and can be readily made by those skilled in the art without departing from the scope and spirit of this invention. Accordingly, it is not intended that the scope of the claims appended hereto be limited to the description as set forth herein, but rather that the claims be broadly construed. [0044]
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1
3
1
4310
DNA
Oryza sativa
CDS
(655)..(3828)
1
ctcttctcca ctccaaatcc cttcttaccc tattcccctc cccccgcagc ttctcttcct 60
cctgcagtac tcgccgccac caccaccgcg ccgccgccgc cggccgcgtt ccgagaccca 120
ctcgatcgga atccaccgcg gcgcgcccgc gcgcctgcgt cctcttcctt ccccgggagc 180
cgaccgacca cggcgaccag tcgatctccc tctccgggcg ccaaccgcgt cttagcttca 240
tcgaatccac cgccccaccc cgcatctcct cctcctcctc cgacgacgac gactactact 300
agtcttctcc aataagcccc cctcccgctc cccccgcctg aagaagaagc agcagctagc 360
tccggggaga ggtcgacggc gcgccgggta gatcgcgccc cgccccgcct gcgtcgcggc 420
tgtcggagca aacgcaaacc ccccaggttg ttctagcgtg tgcagcggct agctgattga 480
ttgtcttctg tgatatatcc agagctcgtg ttttgtggtt tgtggtttgt ggtttgtgct 540
tggattgttg atgtgctaat tcgcggcgtt acaagatcac tgctggattg atattgagtt 600
gtgcctcggc tgtgctagct gtgtgttgat tctctcctcg tcgtggtgat cgat atg 657
Met
1
gag att gtt gca gta gat cag gag gga gct cgt gtt gtt ggg acg aac 705
Glu Ile Val Ala Val Asp Gln Glu Gly Ala Arg Val Val Gly Thr Asn
5 10 15
tgt atg ctt gct cgt ggt gga act ggt gct gta gcg cca gtg ttg gag 753
Cys Met Leu Ala Arg Gly Gly Thr Gly Ala Val Ala Pro Val Leu Glu
20 25 30
ctg aca gcg acg cct cgt cag gat gca gcc gct gaa gct ggt gta gac 801
Leu Thr Ala Thr Pro Arg Gln Asp Ala Ala Ala Glu Ala Gly Val Asp
35 40 45
gaa ccg gca caa cac caa tgc gag cat ttc tcc ata aga ggg tat gtt 849
Glu Pro Ala Gln His Gln Cys Glu His Phe Ser Ile Arg Gly Tyr Val
50 55 60 65
gct ctt ctt cag aag aag gat cca aaa ttc tgc tct cta tct cgg att 897
Ala Leu Leu Gln Lys Lys Asp Pro Lys Phe Cys Ser Leu Ser Arg Ile
70 75 80
ttc cat gac cag aaa aaa tgt gat gaa cac aaa gct agt tca agc cca 945
Phe His Asp Gln Lys Lys Cys Asp Glu His Lys Ala Ser Ser Ser Pro
85 90 95
ttt tct gta gca aag ttt cga cga tgg gat tgc tcg aag tgc ttg gat 993
Phe Ser Val Ala Lys Phe Arg Arg Trp Asp Cys Ser Lys Cys Leu Asp
100 105 110
aag ttg aaa act tca gat aat gga aca gca cca aga act ctt ccc gca 1041
Lys Leu Lys Thr Ser Asp Asn Gly Thr Ala Pro Arg Thr Leu Pro Ala
115 120 125
aag cag aat ggc aca agt gat ggt tgc tcc atc aca ttt gtt cgg agc 1089
Lys Gln Asn Gly Thr Ser Asp Gly Cys Ser Ile Thr Phe Val Arg Ser
130 135 140 145
act ttt gtg cct gct agt gtt ggt tcc caa aaa gtg tct cct agc aca 1137
Thr Phe Val Pro Ala Ser Val Gly Ser Gln Lys Val Ser Pro Ser Thr
150 155 160
caa tca tct caa ggg aag aat gct gat aga tca act ctt cca aag agt 1185
Gln Ser Ser Gln Gly Lys Asn Ala Asp Arg Ser Thr Leu Pro Lys Ser
165 170 175
gtg caa gaa ggc aat gac tcc aaa tgc aat gcg cct tct ggc aag aat 1233
Val Gln Glu Gly Asn Asp Ser Lys Cys Asn Ala Pro Ser Gly Lys Asn
180 185 190
gga gct gct gag gcc aat act gat tca cca atg aaa gat ttg caa ggg 1281
Gly Ala Ala Glu Ala Asn Thr Asp Ser Pro Met Lys Asp Leu Gln Gly
195 200 205
cca gcc caa aat tat gat gtg gca gca aat gtc tct gag gac aac act 1329
Pro Ala Gln Asn Tyr Asp Val Ala Ala Asn Val Ser Glu Asp Asn Thr
210 215 220 225
tct gtt gat gtt ggg gct tta cct gaa gtt ccc cag att aca tgg cac 1377
Ser Val Asp Val Gly Ala Leu Pro Glu Val Pro Gln Ile Thr Trp His
230 235 240
ata gaa gta aat ggt gca gat caa cct cca tcc act cca aaa ctt tct 1425
Ile Glu Val Asn Gly Ala Asp Gln Pro Pro Ser Thr Pro Lys Leu Ser
245 250 255
gaa gtg gtc ctc aaa aga aat gaa gat gaa aat gga aaa act gaa gag 1473
Glu Val Val Leu Lys Arg Asn Glu Asp Glu Asn Gly Lys Thr Glu Glu
260 265 270
act ctt gtt gct gag cag tgc aat ttg acc aaa gat cct aac cca atg 1521
Thr Leu Val Ala Glu Gln Cys Asn Leu Thr Lys Asp Pro Asn Pro Met
275 280 285
tct gga aag gaa cgt gat cag gtt gct gag cag tgc aat ttg acc aaa 1569
Ser Gly Lys Glu Arg Asp Gln Val Ala Glu Gln Cys Asn Leu Thr Lys
290 295 300 305
gat ccg aaa cca gtg tct ggg cag aaa tgt gag cag atc tgc aat gag 1617
Asp Pro Lys Pro Val Ser Gly Gln Lys Cys Glu Gln Ile Cys Asn Glu
310 315 320
cca tgt gaa gaa gtt gtt ctc aaa aga agc tcc aaa tct aag agg aag 1665
Pro Cys Glu Glu Val Val Leu Lys Arg Ser Ser Lys Ser Lys Arg Lys
325 330 335
acg gat aag aag ttg atg aag aag cag cag cac agc aag aaa cgc act 1713
Thr Asp Lys Lys Leu Met Lys Lys Gln Gln His Ser Lys Lys Arg Thr
340 345 350
gcc cag gct gat gtt tca gat gca aag ctt tgt cgg aga aag cca aaa 1761
Ala Gln Ala Asp Val Ser Asp Ala Lys Leu Cys Arg Arg Lys Pro Lys
355 360 365
aag gtg cgg ctt cta tca gaa att ata aat gct aac cag gtt gag gat 1809
Lys Val Arg Leu Leu Ser Glu Ile Ile Asn Ala Asn Gln Val Glu Asp
370 375 380 385
tct aga agt gac gaa gtt cat cgt gaa aat gcc gct gat ccc tgt gag 1857
Ser Arg Ser Asp Glu Val His Arg Glu Asn Ala Ala Asp Pro Cys Glu
390 395 400
gat gat aga agt acc atc ccg gtc ccg atg gaa gta agc atg gat att 1905
Asp Asp Arg Ser Thr Ile Pro Val Pro Met Glu Val Ser Met Asp Ile
405 410 415
cct gtt agc aac cat aca gtg gga gaa gat ggg tta aaa tca agt aag 1953
Pro Val Ser Asn His Thr Val Gly Glu Asp Gly Leu Lys Ser Ser Lys
420 425 430
aac aag aca aaa cgc aaa tac tct gat gtt gta gat gat gga tca tca 2001
Asn Lys Thr Lys Arg Lys Tyr Ser Asp Val Val Asp Asp Gly Ser Ser
435 440 445
ctt atg aac tgg ctg aat gga aaa aag aaa aga act gga agt gtg cat 2049
Leu Met Asn Trp Leu Asn Gly Lys Lys Lys Arg Thr Gly Ser Val His
450 455 460 465
cac aca gtt gct cat cca gct ggg aat ttg agc aac aaa aaa gtg aca 2097
His Thr Val Ala His Pro Ala Gly Asn Leu Ser Asn Lys Lys Val Thr
470 475 480
ccc act gcg agt act cag cat gat gat gag aat gat act gaa aat ggt 2145
Pro Thr Ala Ser Thr Gln His Asp Asp Glu Asn Asp Thr Glu Asn Gly
485 490 495
ctt gac aca aat atg cat aag aca gat gtc tgt cag cat gta tca gaa 2193
Leu Asp Thr Asn Met His Lys Thr Asp Val Cys Gln His Val Ser Glu
500 505 510
atc tcc aca cag agg tgc tca tca aag ggg aaa aca gcg ggt ttg agt 2241
Ile Ser Thr Gln Arg Cys Ser Ser Lys Gly Lys Thr Ala Gly Leu Ser
515 520 525
aag ggg aaa aca cat tca gct gct agt acc aaa tat ggt ggt gaa agc 2289
Lys Gly Lys Thr His Ser Ala Ala Ser Thr Lys Tyr Gly Gly Glu Ser
530 535 540 545
acc aga aat ggt cag aac ata cat gta ctc agc gca gaa gat caa tgc 2337
Thr Arg Asn Gly Gln Asn Ile His Val Leu Ser Ala Glu Asp Gln Cys
550 555 560
cag atg gaa acc gaa aac tct gtt ctg agt cac tcg gca aag gtt tct 2385
Gln Met Glu Thr Glu Asn Ser Val Leu Ser His Ser Ala Lys Val Ser
565 570 575
cca gct gag cat gat atc caa att atg tct gac ctt cat gag cag agt 2433
Pro Ala Glu His Asp Ile Gln Ile Met Ser Asp Leu His Glu Gln Ser
580 585 590
cta ccc aag aag aaa aag aag caa aaa ctt gaa gtg act cgt gaa aaa 2481
Leu Pro Lys Lys Lys Lys Lys Gln Lys Leu Glu Val Thr Arg Glu Lys
595 600 605
cag acc atg ata gat gac atc ccc atg gat att gtt gaa ctg cta gct 2529
Gln Thr Met Ile Asp Asp Ile Pro Met Asp Ile Val Glu Leu Leu Ala
610 615 620 625
aaa aac cag cat gag agg cag ctt atg act gag act gat tgt tct gac 2577
Lys Asn Gln His Glu Arg Gln Leu Met Thr Glu Thr Asp Cys Ser Asp
630 635 640
atc aac cgt att caa tcc aag aca act gct gat gat gat tgt gta ata 2625
Ile Asn Arg Ile Gln Ser Lys Thr Thr Ala Asp Asp Asp Cys Val Ile
645 650 655
gta gct gcc aag gat ggt tca gat tat gca tca agt gtg ttt gac act 2673
Val Ala Ala Lys Asp Gly Ser Asp Tyr Ala Ser Ser Val Phe Asp Thr
660 665 670
aat tcc caa cag aag tcc ttg gca tcc caa agt aca cag aag gag tta 2721
Asn Ser Gln Gln Lys Ser Leu Ala Ser Gln Ser Thr Gln Lys Glu Leu
675 680 685
cag ggt cat ttg gca ttg acc aca caa gag tct cca cat cct cag aac 2769
Gln Gly His Leu Ala Leu Thr Thr Gln Glu Ser Pro His Pro Gln Asn
690 695 700 705
ttt cag tct act cag gaa cag cag aca cat ttg cgg atg gaa gaa atg 2817
Phe Gln Ser Thr Gln Glu Gln Gln Thr His Leu Arg Met Glu Glu Met
710 715 720
gtc act att gct gca agc tca cca cta ttt tca cat cat gat gat cag 2865
Val Thr Ile Ala Ala Ser Ser Pro Leu Phe Ser His His Asp Asp Gln
725 730 735
tat att gct gaa gca cca act gaa cat tgg ggc cgt aag gac gca aag 2913
Tyr Ile Ala Glu Ala Pro Thr Glu His Trp Gly Arg Lys Asp Ala Lys
740 745 750
aag cta acg tgg gag caa ttt aag gcc act aca aga aat tct cca gca 2961
Lys Leu Thr Trp Glu Gln Phe Lys Ala Thr Thr Arg Asn Ser Pro Ala
755 760 765
gca aca tgt ggt gct caa ttt aga cct ggt atc caa gca gtt gac ttg 3009
Ala Thr Cys Gly Ala Gln Phe Arg Pro Gly Ile Gln Ala Val Asp Leu
770 775 780 785
act tct act cat gtc atg gga tct tcc agc aat tat gca tct cgc caa 3057
Thr Ser Thr His Val Met Gly Ser Ser Ser Asn Tyr Ala Ser Arg Gln
790 795 800
cca gta att gcg cca ctg gac cgc tat gct gaa aga gcg gtt aac cag 3105
Pro Val Ile Ala Pro Leu Asp Arg Tyr Ala Glu Arg Ala Val Asn Gln
805 810 815
gtc cat gca aga aat ttt cca agc aca ata gca acc atg gaa gcg agt 3153
Val His Ala Arg Asn Phe Pro Ser Thr Ile Ala Thr Met Glu Ala Ser
820 825 830
aag tta tgt gat cgg aga aat gct gga caa gta gtc ttg tat cct aaa 3201
Lys Leu Cys Asp Arg Arg Asn Ala Gly Gln Val Val Leu Tyr Pro Lys
835 840 845
gaa tcc atg cct gcg acg cat ctt ctg aga atg atg gat cca tca aca 3249
Glu Ser Met Pro Ala Thr His Leu Leu Arg Met Met Asp Pro Ser Thr
850 855 860 865
tta gca agc ttc ccc aac tat gga act tct agc agg aac cag atg gag 3297
Leu Ala Ser Phe Pro Asn Tyr Gly Thr Ser Ser Arg Asn Gln Met Glu
870 875 880
tct caa ctt cat aat tct cag tat gca cat aat cag tac aaa gga tca 3345
Ser Gln Leu His Asn Ser Gln Tyr Ala His Asn Gln Tyr Lys Gly Ser
885 890 895
acc agc aca tca tat ggc agt aac ctg aat gga aag att cca ttg aca 3393
Thr Ser Thr Ser Tyr Gly Ser Asn Leu Asn Gly Lys Ile Pro Leu Thr
900 905 910
ttc gaa gac tta tca cgg cat cag ctg cat gat ctg cac aga cct tta 3441
Phe Glu Asp Leu Ser Arg His Gln Leu His Asp Leu His Arg Pro Leu
915 920 925
cgc cca cat cct aga gtt ggt gtg ctt ggc tcc ttg ctg cag aag gaa 3489
Arg Pro His Pro Arg Val Gly Val Leu Gly Ser Leu Leu Gln Lys Glu
930 935 940 945
att gca aac tgg tcg gag aac tgt ggc aca caa tct ggt tat aag tta 3537
Ile Ala Asn Trp Ser Glu Asn Cys Gly Thr Gln Ser Gly Tyr Lys Leu
950 955 960
gga gtg tca aca gga ata aca tcg cat cag atg aac aga aag gaa cat 3585
Gly Val Ser Thr Gly Ile Thr Ser His Gln Met Asn Arg Lys Glu His
965 970 975
ttt gaa gcc ctg aat tct gga atg ttt tca gca aaa tgg aat gca ttg 3633
Phe Glu Ala Leu Asn Ser Gly Met Phe Ser Ala Lys Trp Asn Ala Leu
980 985 990
cag ttg ggt tct gtt agc tcc agt gca gat ttt tta tca gcg agg aac 3681
Gln Leu Gly Ser Val Ser Ser Ser Ala Asp Phe Leu Ser Ala Arg Asn
995 1000 1005
agc ata gct caa tct tgg acc aga ggc aag ggt aaa atg gtt cat ccc 3729
Ser Ile Ala Gln Ser Trp Thr Arg Gly Lys Gly Lys Met Val His Pro
1010 1015 1020 1025
ttg gat cgg ttt gtg aga cag gat atc tgt ata act aac aag aac cca 3777
Leu Asp Arg Phe Val Arg Gln Asp Ile Cys Ile Thr Asn Lys Asn Pro
1030 1035 1040
gct gat ttt act aca atc agt aac gat aac gag tat atg gat tac cgc 3825
Ala Asp Phe Thr Thr Ile Ser Asn Asp Asn Glu Tyr Met Asp Tyr Arg
1045 1050 1055
tga agcagaaagt ggtgtgcata attcctgaac atttacaatc atacatttca 3878
tctttatggc gccaaatagt catactgtaa gaggagggct ttgctggatc tgctgtaagg 3938
cttcttgtaa gttgtggatg ccccattttc tggatgggaa cctgccagac agtgaacaag 3998
ggctttgcaa ggtgcagcat ccggtttttg ttttgccagt ccaagaaacg tcctcctgtt 4058
actttgtagt tgtactcata ctagtgcgct tgtttgtaca aggagaaatg tgtaaccttg 4118
ttgaaaaaat gtctccccca ttttgtaatt accataagga ggtttatagt gttgtgagct 4178
gtgtgtgact gacggcgaga aatggttttg tcggtgttaa ggttgaaacg actagctctc 4238
gttatcaatg tgttgtaaac ttctagattg atgtgttacc ttactcttga agtcaacacc 4298
ggagaattta ca 4310
2
1057
PRT
Oryza sativa
2
Met Glu Ile Val Ala Val Asp Gln Glu Gly Ala Arg Val Val Gly Thr
1 5 10 15
Asn Cys Met Leu Ala Arg Gly Gly Thr Gly Ala Val Ala Pro Val Leu
20 25 30
Glu Leu Thr Ala Thr Pro Arg Gln Asp Ala Ala Ala Glu Ala Gly Val
35 40 45
Asp Glu Pro Ala Gln His Gln Cys Glu His Phe Ser Ile Arg Gly Tyr
50 55 60
Val Ala Leu Leu Gln Lys Lys Asp Pro Lys Phe Cys Ser Leu Ser Arg
65 70 75 80
Ile Phe His Asp Gln Lys Lys Cys Asp Glu His Lys Ala Ser Ser Ser
85 90 95
Pro Phe Ser Val Ala Lys Phe Arg Arg Trp Asp Cys Ser Lys Cys Leu
100 105 110
Asp Lys Leu Lys Thr Ser Asp Asn Gly Thr Ala Pro Arg Thr Leu Pro
115 120 125
Ala Lys Gln Asn Gly Thr Ser Asp Gly Cys Ser Ile Thr Phe Val Arg
130 135 140
Ser Thr Phe Val Pro Ala Ser Val Gly Ser Gln Lys Val Ser Pro Ser
145 150 155 160
Thr Gln Ser Ser Gln Gly Lys Asn Ala Asp Arg Ser Thr Leu Pro Lys
165 170 175
Ser Val Gln Glu Gly Asn Asp Ser Lys Cys Asn Ala Pro Ser Gly Lys
180 185 190
Asn Gly Ala Ala Glu Ala Asn Thr Asp Ser Pro Met Lys Asp Leu Gln
195 200 205
Gly Pro Ala Gln Asn Tyr Asp Val Ala Ala Asn Val Ser Glu Asp Asn
210 215 220
Thr Ser Val Asp Val Gly Ala Leu Pro Glu Val Pro Gln Ile Thr Trp
225 230 235 240
His Ile Glu Val Asn Gly Ala Asp Gln Pro Pro Ser Thr Pro Lys Leu
245 250 255
Ser Glu Val Val Leu Lys Arg Asn Glu Asp Glu Asn Gly Lys Thr Glu
260 265 270
Glu Thr Leu Val Ala Glu Gln Cys Asn Leu Thr Lys Asp Pro Asn Pro
275 280 285
Met Ser Gly Lys Glu Arg Asp Gln Val Ala Glu Gln Cys Asn Leu Thr
290 295 300
Lys Asp Pro Lys Pro Val Ser Gly Gln Lys Cys Glu Gln Ile Cys Asn
305 310 315 320
Glu Pro Cys Glu Glu Val Val Leu Lys Arg Ser Ser Lys Ser Lys Arg
325 330 335
Lys Thr Asp Lys Lys Leu Met Lys Lys Gln Gln His Ser Lys Lys Arg
340 345 350
Thr Ala Gln Ala Asp Val Ser Asp Ala Lys Leu Cys Arg Arg Lys Pro
355 360 365
Lys Lys Val Arg Leu Leu Ser Glu Ile Ile Asn Ala Asn Gln Val Glu
370 375 380
Asp Ser Arg Ser Asp Glu Val His Arg Glu Asn Ala Ala Asp Pro Cys
385 390 395 400
Glu Asp Asp Arg Ser Thr Ile Pro Val Pro Met Glu Val Ser Met Asp
405 410 415
Ile Pro Val Ser Asn His Thr Val Gly Glu Asp Gly Leu Lys Ser Ser
420 425 430
Lys Asn Lys Thr Lys Arg Lys Tyr Ser Asp Val Val Asp Asp Gly Ser
435 440 445
Ser Leu Met Asn Trp Leu Asn Gly Lys Lys Lys Arg Thr Gly Ser Val
450 455 460
His His Thr Val Ala His Pro Ala Gly Asn Leu Ser Asn Lys Lys Val
465 470 475 480
Thr Pro Thr Ala Ser Thr Gln His Asp Asp Glu Asn Asp Thr Glu Asn
485 490 495
Gly Leu Asp Thr Asn Met His Lys Thr Asp Val Cys Gln His Val Ser
500 505 510
Glu Ile Ser Thr Gln Arg Cys Ser Ser Lys Gly Lys Thr Ala Gly Leu
515 520 525
Ser Lys Gly Lys Thr His Ser Ala Ala Ser Thr Lys Tyr Gly Gly Glu
530 535 540
Ser Thr Arg Asn Gly Gln Asn Ile His Val Leu Ser Ala Glu Asp Gln
545 550 555 560
Cys Gln Met Glu Thr Glu Asn Ser Val Leu Ser His Ser Ala Lys Val
565 570 575
Ser Pro Ala Glu His Asp Ile Gln Ile Met Ser Asp Leu His Glu Gln
580 585 590
Ser Leu Pro Lys Lys Lys Lys Lys Gln Lys Leu Glu Val Thr Arg Glu
595 600 605
Lys Gln Thr Met Ile Asp Asp Ile Pro Met Asp Ile Val Glu Leu Leu
610 615 620
Ala Lys Asn Gln His Glu Arg Gln Leu Met Thr Glu Thr Asp Cys Ser
625 630 635 640
Asp Ile Asn Arg Ile Gln Ser Lys Thr Thr Ala Asp Asp Asp Cys Val
645 650 655
Ile Val Ala Ala Lys Asp Gly Ser Asp Tyr Ala Ser Ser Val Phe Asp
660 665 670
Thr Asn Ser Gln Gln Lys Ser Leu Ala Ser Gln Ser Thr Gln Lys Glu
675 680 685
Leu Gln Gly His Leu Ala Leu Thr Thr Gln Glu Ser Pro His Pro Gln
690 695 700
Asn Phe Gln Ser Thr Gln Glu Gln Gln Thr His Leu Arg Met Glu Glu
705 710 715 720
Met Val Thr Ile Ala Ala Ser Ser Pro Leu Phe Ser His His Asp Asp
725 730 735
Gln Tyr Ile Ala Glu Ala Pro Thr Glu His Trp Gly Arg Lys Asp Ala
740 745 750
Lys Lys Leu Thr Trp Glu Gln Phe Lys Ala Thr Thr Arg Asn Ser Pro
755 760 765
Ala Ala Thr Cys Gly Ala Gln Phe Arg Pro Gly Ile Gln Ala Val Asp
770 775 780
Leu Thr Ser Thr His Val Met Gly Ser Ser Ser Asn Tyr Ala Ser Arg
785 790 795 800
Gln Pro Val Ile Ala Pro Leu Asp Arg Tyr Ala Glu Arg Ala Val Asn
805 810 815
Gln Val His Ala Arg Asn Phe Pro Ser Thr Ile Ala Thr Met Glu Ala
820 825 830
Ser Lys Leu Cys Asp Arg Arg Asn Ala Gly Gln Val Val Leu Tyr Pro
835 840 845
Lys Glu Ser Met Pro Ala Thr His Leu Leu Arg Met Met Asp Pro Ser
850 855 860
Thr Leu Ala Ser Phe Pro Asn Tyr Gly Thr Ser Ser Arg Asn Gln Met
865 870 875 880
Glu Ser Gln Leu His Asn Ser Gln Tyr Ala His Asn Gln Tyr Lys Gly
885 890 895
Ser Thr Ser Thr Ser Tyr Gly Ser Asn Leu Asn Gly Lys Ile Pro Leu
900 905 910
Thr Phe Glu Asp Leu Ser Arg His Gln Leu His Asp Leu His Arg Pro
915 920 925
Leu Arg Pro His Pro Arg Val Gly Val Leu Gly Ser Leu Leu Gln Lys
930 935 940
Glu Ile Ala Asn Trp Ser Glu Asn Cys Gly Thr Gln Ser Gly Tyr Lys
945 950 955 960
Leu Gly Val Ser Thr Gly Ile Thr Ser His Gln Met Asn Arg Lys Glu
965 970 975
His Phe Glu Ala Leu Asn Ser Gly Met Phe Ser Ala Lys Trp Asn Ala
980 985 990
Leu Gln Leu Gly Ser Val Ser Ser Ser Ala Asp Phe Leu Ser Ala Arg
995 1000 1005
Asn Ser Ile Ala Gln Ser Trp Thr Arg Gly Lys Gly Lys Met Val His
1010 1015 1020
Pro Leu Asp Arg Phe Val Arg Gln Asp Ile Cys Ile Thr Asn Lys Asn
1025 1030 1035 1040
Pro Ala Asp Phe Thr Thr Ile Ser Asn Asp Asn Glu Tyr Met Asp Tyr
1045 1050 1055
Arg
3
9455
DNA
Oryza sativa
3
gatctatcgt tgattggatt tcgctgggct agctaggtag acaagggttc aaaatgtgac 60
gaacattatt gagctaataa ataacgagaa accaacttgg tatataggtc aatttcaaaa 120
gaaacaagct gacaaaattc gtccaatttc actagttttt gtcagtaatt gaatggcaat 180
catggttatc gacaaaaccg cttaggagtg ctatttggtg atggaatggt ttataaaact 240
ttggaccgga gtagcagtac aatggcttgt ctgaacaggc taggtagcat agtaggtcct 300
tttgccttgg ttgcactgtt ctgtcggctt ataggggaat ctattggctt aatggaaggg 360
aaaatagtgt gactagcatc atcgatttgc ttgctatcat gtttgagcat cattgacatg 420
tgggtgtcga tctaggagac tatgaatcta gcgaatcctc actagtatgc acatgcacaa 480
cgacatcatg cagctatatg tacaacaatg taggaatggt agctctactc ggattggtgg 540
ccgcattcct acattgctag aaaaacacaa acacacacac acacacacac aaacaaacat 600
aggacgctgc aacacctctt ccctggctac cactatcgcc aaatgccaag catggttgtg 660
ttgacctccc tccttagctg ccactgtgtc cacttcaaaa tccataacca cgacatcttc 720
ccttcttcta caattgttac tactgttctt gaagaggttc ttgtcctacc gacttactaa 780
agaccaacca ttggtcagca gagcatctcg agacttctat accaaagtaa cattgagagg 840
tcaatggcat gtaggaccat cactccgttt gcattgacca atcagagatg agtctgaaca 900
atttcaaccc gtaaaaacgg tccacaattt gattcatcta ttggtttgct gtctgcccat 960
ggccctgcat cttgattgtt tatgatcttt cttagttatc tcttgagatt taatctacct 1020
aattgtgtca ttttaatcct ttgatttatc ttgattggat tcggtctact ttggtttcac 1080
catcttagcc ccgatatgat cttggtagcc cttttttcct cgcactaatc attttcgatt 1140
ctcgactcca atctaagttc ccatggtaga tttaggtcaa taagtgaaaa acaatttccc 1200
ttacaattta atcccctttt gagtaatgac cttcgtactt cctcaaacga tggtttcatg 1260
gataaagatt tgatatttgt ggcgcacgga tggggcatgg cttatgaaga aaatgagctc 1320
attggattgg tattttctcc tccatgctcc tattggtaaa aatattggtt tagttggata 1380
gtcaggttag aacgggattt aaaaatgtga ctacaactag tcgtttatcg cggttgcaca 1440
tgggtgtcaa cgggtgactg cctgtgttgg catcaataat ttagaattag aggtgtcggc 1500
ttaatgctgg tagattgtgg atatcctccc atacacgtag gcgtcgatgg cttcatgtgc 1560
gtacgtgccg tcctgcatag gatgggtagc aacggaggaa ggctggcttt gttgacttgt 1620
agcctcacta aggccaactg cctattggca atgaagttgt ttttgtttaa ggagaaacaa 1680
acttttgagc taggcaacat cggtatgtag caggccattc tctaggtcaa tggcacttag 1740
agtcggagca tgccggccgt gcctattaca acatggttat catgtgcatg tggcaagggc 1800
acacattaca cgtccccatt gtctaacatc gtagactatg agtttgatag aagaaaccca 1860
actgaaaggt cagtgtcaaa tccattctac ctgtgtcgag aactaaggtt aagcgtttta 1920
tatagattag gacactcaca tacatatgct gtgcatgggg tggtgagagc cattatgtac 1980
cactagtgac atgtgtctac ctagttatat gatctattga ttgaaaattc gactgagata 2040
agaataacta tgtggtatat ttaacttggt acaacatata gtggttggaa catcacatta 2100
gacatgcctt ggcaggaaga aagccgctgt agtttaattt aacatgttaa acaccacata 2160
gtgagtaatc aatctaggct taaaaaaaat aaagacaaca caaacaaaca cagagaacac 2220
ataaacacag agggggcaat caactctgtt gtttgatcta tacttcgtca gagtttgata 2280
tatctctgct ctctgcattt taaaactact tatattacaa agcactgtat gttatatagt 2340
aatactttgt agatcacgag tataaaacac agataggtac aatttttttt actcctacgt 2400
atttacgtat taaccggtac aatattattt agcatgcatg tacgcgtgat attagagtgg 2460
agtatactac ctacgcatgc tcgggcagta tcggtacctc tatacctacc tatatacaca 2520
taccaggtag agtacagcgg caggtagagt acacagtagt atatggagct tattttatca 2580
tttttattac tggagataga agaagaagag aaaggagaga gagagagaaa gagtggaagg 2640
gaaaggcgtg cggggcccac acacgcagca gggcagcgct gcggaggtga ggtgagggca 2700
ggggcagggg cagggcggac tggccaacgc caactccaac tccaactccc gtacaaataa 2760
aatatacaat cctttctttt tcctcctcct cttttccctt ccattccacc cccctctctc 2820
tctcttctcc actccaaatc ccttcttacc ctattcccct ccccccgcag cttctcttcc 2880
tcctgcagta ctcgccgcca ccaccaccgc gccgccgccg ccggccgcgt tccgagaccc 2940
actcgatcgg aatccaccgc ggcgcgcccg cgcgcctgcg tcctcttcct tccccgggag 3000
ccgaccgacc acggcgacca gtcgatctcc ctctccgggc gccaaccgcg tcttagcttc 3060
atcgaatcca ccgccccacc ccgcatctcc tcctcctcct ccgacgacga cgactactac 3120
tagtcttctc caataagccc ccctcccgct ccccccgcct gaagaagaag cagcagctag 3180
ctccggggag aggtcgacgg cgcgccgggt agatcgcgcc ccgccccgcc tgcgtcgcgg 3240
ctgtcggagc aaacgcaaac cccccaggta atcaacgaac ttttcctccg ccgcaagaac 3300
agctcccgcg gggggtttgg ttttgaccga tttcttcccc cctcccccca aatcgaccca 3360
tccaatttcg cctcgattta cttccgattt ccccactttt ttttcttcct ttcgggttgg 3420
ggggttgcgg ttttggggga ggagaggggt tcagctcatc cgaagcccca cgttaggtcc 3480
gccccctttc cagctgtgcc cctctctcgg gcctcgagct cctcgcctcc atgggaacca 3540
aagcccttat atttcatgtc gcggaagaaa aaaaaatccc gtcttttgcg gggatcctcg 3600
cggctacgta cgagccctag ttaccgcgcg gattttagtt acggcggttt atgcggcccc 3660
tccctctagg ttttagatct acccatctct ctctctctct ctctctctct ctctgtgcat 3720
gcatgtgtct atcttagcta tacctgtatt atttggaagg ttaattatgg ttgtgtatat 3780
gtggcgcggt aattaattag tttaattcgc accccctctc tctttgttta tctaggtttt 3840
gggggaattt atttcttgct ataattttgc ccgctcgaat ttctggtgct cttatattcc 3900
atgagctgat tgaagtggat atatattgtg cgtgcgtgcg tgctattgct acatcggctt 3960
gacttcttct tgcctactac ttcattaatt tgtttcttct ggtttctgtt tcaggttgtt 4020
ctagcgtgtg cagcggctag ctgattgatt gtcttctgtg atatatccag agctcgtgtt 4080
ttgtggtttg tggtttgtgg tttgtgcttg gattgttgat gtgctaattc gcggcgttac 4140
aagatcactg ctggattgat attgagttgt gcctcggctg tgctagctgt gtgttgattc 4200
tctcctcgtc gtggtgatcg atatggagat tgttgcagta gatcaggagg gagctcgtgt 4260
tgttgggacg aactgtatgc ttgctcgtgg tggaactggt gctgtagcgc cagtgttgga 4320
gctgacagcg acgcctcgtc aggatgcagc cgctgaagct ggtgtagacg aaccggcaca 4380
acaccaatgc gagcatttct ccataaggta atcattttct gtatttccaa ttccagtatc 4440
gcgttgtgga tgaataatga atcggcatgt catgccatat tgcactgttt gatggaagag 4500
tatgattgat cgtggttttt gcacagtttg ctgttgggac ttatatggtc atctgttttg 4560
tacgatcgta tacactgggt cgacatgctt atgactttgg ttcgatttag gaagtcaata 4620
catccactac tagctctata tctagccatg tgaactcatt tatgccatag cacagctagc 4680
aggctagcag caaaaaatat atataatatt tgcatatatg ttggtgtttc atgtatcttt 4740
atactctacg tacatccatt aatatcttca atgtatgaat ctgagcacat gattgtgagt 4800
gctacacata tgcatgtctg tatgtgtgtt cattaggtgt ttgatcatat ttgtttgtgt 4860
tggggtgcgc atgcatttat tcaggccatg ctgtaggctg tagctagata tttgtgtttg 4920
tatattattt ctgttgaaca agctgattac taatgaaatt aacctttttg gggtacactc 4980
atatattggg ccctacattt ttgtaatcat ttttcctttg tgctgaggtt cagcataaaa 5040
cttttttatc ataagcatgt ttacatccta ggagattctt agaactgatg gtttcttcat 5100
atttgcatta tgtttgattt gatagtccat tattatttta agccttttca attgtttaga 5160
gattctagag atgatatata tcaaccatag acttgtcacg ttttggttta atactttcta 5220
gaactaatta gattattatt tttgtagttt atcctgtcat gctatttgta ttatctttga 5280
attcaaactg caatacttag attatcttga aggtcctctt tttctggact gtacaagcta 5340
tgtatgaaat gcctacctcc cagcatcctt tagattatgt agggcctttt ctgagtttat 5400
cagttgtata ttgactgaag cacgcaatgt gctatatata tgtgccatgc atctttataa 5460
tgataatctt atttttcttg taccagaggg tatgttgctc ttcttcagaa gaaggatcca 5520
aaattctgct ctctatctcg gattttccat gaccagaaaa aatgtgatga acacaaagct 5580
agttcaagcc cattttctgt agcaaagttt cgacgatggg attgctcgaa gtgcttggat 5640
aagttgaaaa cttcagataa tggaacagca ccaagaactc ttcccgcaaa gcagaatggc 5700
acaagtgatg gttgctccat cacatttgtt cggagcactt ttgtgcctgc tagtgttggt 5760
tcccaaaaag tgtctcctag cacacaatca tctcaaggga agaatgctga tagatcaact 5820
cttccaaaga gtgtgcaaga aggcaatgac tccaaatgca atgcgccttc tggcaagaat 5880
ggagctgctg aggccaatac tgattcacca atgaaaggta tggtagatgt agagcctttc 5940
aaattcctaa gtaggatttt atttaaggta tagaataaac taatgtttgt gtgattttct 6000
cagatttgca agggccagcc caaaattatg atgtggcagc aaatgtctct gaggacaaca 6060
cttctgttga tgttggggct ttacctgaag ttccccagat tacatggcac atagaagtaa 6120
atggtgcaga tcaacctcca tccactccaa aactttctga agtggtcctc aaaagaaatg 6180
aagatgaaaa tggaaaaact gaagagactc ttgttgctga gcagtgcaat ttgaccaaag 6240
atcctaaccc aatgtctgga aaggaacgtg atcaggttgc tgagcagtgc aatttgacca 6300
aagatccgaa accagtgtct gggcagaaat gtgagcagat ctgcaatgag ccatgtgaag 6360
aagttgttct caaaagaagc tccaaatcta agaggaagac ggataagaag ttgatgaaga 6420
agcagcagca cagcaagaaa cgcactgccc aggctgatgt ttcagatgca aagctttgtc 6480
ggagaaagcc aaaaaaggtg cggcttctat cagaaattat aaatgctaac caggttgagg 6540
attctagaag tgacgaagtt catcgtgaaa atgccgctga tccctgtgag gatgatagaa 6600
gtaccatccc ggtcccgatg gaagtaagca tggatattcc tgttagcaac catacagtgg 6660
gagaagatgg gttaaaatca agtaagaaca agacaaaacg caaatactct gatgttgtag 6720
atgatggatc atcacttatg aactggctga atggaaaaaa gaaaagaact ggaagtgtgc 6780
atcacacagt tgctcatcca gctgggaatt tgagcaacaa aaaagtgaca cccactgcga 6840
gtactcagca tgatgatgag aatgatactg aaaatggtct tgacacaaat atgcataaga 6900
cagatgtctg tcagcatgta tcagaaatct ccacacagag gtgctcatca aaggggaaaa 6960
cagcgggttt gagtaagggg aaaacacatt cagctgctag taccaaatat ggtggtgaaa 7020
gcaccagaaa tggtcagaac atacatgtac tcagcgcaga agatcaatgc cagatggaaa 7080
ccgaaaactc tgttctgagt cactcggcaa aggtacgaat tttgtgaatc atgaggaatt 7140
tttgcttttt aaattgactg aatcaacatt tatctgtatg aaggaataat attggtgcat 7200
aacaatgtta agaaatatgc atacaatgtt tatttatatg ctttccactg ttcttcttta 7260
cttatgtttt gatactcttt ttgtgtgtgc gtgcatgtgt gcatgtgtgt gtgtgtgtgt 7320
gtgtgcgcgc gtgtgtgtgc acgtgcgtgg cgcaatattc ttttttagac tcatattata 7380
gtgattgtaa tggactgaca ttttcctcat ttctcatctc aggtttctcc agctgagcat 7440
gatatccaaa ttatgtctga ccttcatgag cagagtctac ccaagaagaa aaagaagcaa 7500
aaacttgaag tgactcgtga aaaacagacc atgatagatg acatccccat ggatattgtt 7560
gaactgctag ctaaaaacca gcatgagagg cagcttatga ctgagactga ttgttctgac 7620
atcaaccgta ttcaatccaa gacaactgct gatgatgatt gtgtaatagt agctgccaag 7680
gatggttcag attatgcatc aagtgtgttt gacactaatt cccaacagaa gtccttggca 7740
tcccaaagta cacagaagga gttacagggt catttggcat tgaccacaca agagtctcca 7800
catcctcaga actttcagtc tactcaggaa cagcagacac atttgcggat ggaagaaatg 7860
gtcactattg ctgcaagctc accactattt tcacatcatg atgatcagta tattgctgaa 7920
gcaccaactg aacattgggg ccgtaaggac gcaaagaagc taacgtggga gcaatttaag 7980
gccactacaa gaaattctcc agcagcaaca tgtggtgctc aatttagacc tggtatccaa 8040
gcagttgact tgacttctac tcatgtcatg ggatcttcca gcaattatgc atctcgccaa 8100
ccagtaattg cgccactgga ccgctatgct gaaagagcgg ttaaccaggt ccatgcaaga 8160
aattttccaa gcacaatagc aaccatggaa gcgagtaagt tatgtgatcg gagaaatgct 8220
ggacaagtag tcttgtatcc taaagaatcc atgcctgcga cgcatcttct gagaatgatg 8280
gatccatcaa cattagcaag cttccccaac tatggaactt ctagcaggaa ccagatggag 8340
tctcaacttc ataattctca gtatgcacat aatcagtaca aaggatcaac cagcacatca 8400
tatggcagta acctgaatgg aaagattcca ttgacattcg aagacttatc acggcatcag 8460
ctgcatgatc tgcacagacc tttacgccca catcctagag ttggtgtgct tggctccttg 8520
ctgcagaagg aaattgcaaa ctggtcggag aactgtggca cacaatctgg ttataagtta 8580
ggagtgtcaa caggaataac atcgcatcag atgaacagaa aggaacattt tgaagccctg 8640
aattctggaa tgttttcagc aaaatggaat gcattgcagt tgggttctgt tagctccagt 8700
gcagattttt tatcagcgag gaacagcata gctcaatctt ggaccagagg caagggtaaa 8760
atggttcatc ccttggatcg gtttgtgaga caggatatct gtataactaa caagaaccca 8820
gctgatttta ctacaatcag taacgataac gagtatatgg attaccgctg aagcagaaag 8880
tggtgtgcat aattcctgaa catttacaat catacatttc atctttatgg cgccaaatag 8940
tcatactgta agaggagggc tttgctggat ctgctgtaag gtaagttgaa ctttttcttc 9000
ttgcaagttt atcagtttaa gaaaaaagaa tgattactta tgttagcaag gatggttctt 9060
gcaggcttct tgtaagttgt ggatgcccca ttttctggat gggaacctgc cagacagtga 9120
acaagggctt tgcaaggtgc agcatccggt ttttgttttg ccagtccaag aaacgtcctc 9180
ctgttacttt gtagttgtac tcatactagt gcgcttgttt gtacaaggag aaatgtgtaa 9240
ccttgttgaa aaaatgtctc ccccattttg taattaccat aaggaggttt atagtgttgt 9300
gagctgtgtg tgactgacgg cgagaaatgg ttttgtcggt gttaaggttg aaacgactag 9360
ctctcgttat caatgtgttg taaacttcta gattgatgtg ttaccttact cttgaagtca 9420
acaccggaga atttacagta cttttttgcc gtgcc 9455